应用草履虫建立中药快速毒性筛选实验方法的探讨
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摘要
应用草履虫进行中药快速毒性实验,拟建立药物的单细胞动物快速毒性筛选实验的基本方法.分别进行慢性与急性毒性实验观察草履虫的应激性、运动状态、存活只数.结果显示,在慢性实验的48 h内,草履虫的存活数量与时间、质量浓度呈指数递增.当中药质量浓度为5 mg·mL~(-1)时,草履虫开始出现死亡.急性毒性实验6 h内,药物低质量浓度组未见草履虫死亡.而中、高质量浓度组,随着药物质量浓度、时间增加,毒性也增加.上述研究结果表明,急性毒性实验可快速获得药物安全剂量、最大耐受剂量、致死剂量、半数致死量等参数,可快速进行药物毒性评价和毒性药物筛选.
The research aimed to set up a rapid method for acute toxicity test of traditional Chinese medicine using single cell animal model. Chronic and acute toxicity experiments were conducted to observe the stress,movement,and survival of Paramecium caudatum.The results showed that the survival of paramecium increased exponentially with time and concentration in the 48 h chronic experiment.When the concentration of medicine was 5 mg·mL~(-1),paramecium began to die.In acute toxicity experiment 6 h,no death of paramecium was observed in low concentration group.In the middle and high concentration group,the toxicity increased with the increase of drug concentration and time.The results of the above studies show that acute toxicity experiments can quickly obtain parameters such as drug safety dose,maximum tolerable dose,lethal dose,and median lethal dose,and it can quickly perform drug toxicity evaluation and toxic drug screening.
The research aimed to set up a rapid method for acute toxicity test of traditional Chinese medicine using single cell animal model. Chronic and acute toxicity experiments were conducted to observe the stress,movement,and survival of Paramecium caudatum.The results showed that the survival of paramecium increased exponentially with time and concentration in the 48 h chronic experiment.When the concentration of medicine was 5 mg·mL~(-1),paramecium began to die.In acute toxicity experiment 6 h,no death of paramecium was observed in low concentration group.In the middle and high concentration group,the toxicity increased with the increase of drug concentration and time.The results of the above studies show that acute toxicity experiments can quickly obtain parameters such as drug safety dose,maximum tolerable dose,lethal dose,and median lethal dose,and it can quickly perform drug toxicity evaluation and toxic drug screening.
引文
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[14]RICHARD M,JAMES W,ANDREW A.Toxicity and applications of internalised magnetite nanoparticles within live paramecium caudatum cells[J].Bio Nano Science,2018,8(1):90-94.
[15]MORGUNOV I G,KARPUKHINA O V,KAMZOLOVA S V,et al.Investigation of the effect of biologically active threo-Ds-isocitric acid on oxidative stress in Paramecium caudatum[J].Prep Biochem Biotechnol,2018,48(1):1-5.
[2]张立军,李芸,张艳霞,等.高乌头不同溶剂萃取部分对小鼠的急性毒性研究[J].中国中医药信息杂志,2018,25(3):53-57.
[3]BURATTI F M,SCARDALA S,FUNARI E,et al.The conjugation of microcystin-RR by human recombinant GSTs and hepatic cytosol[J].Toxicol Lett,2013,219(3):231-238.
[4]MANSANO A S,MOREIRA R A,PIEROZZI M,et al.Effects of diuron and carbofuran pesticides in their pure and commercial forms on Paramecium caudatum:the use of protozoan in ecotoxicology [J].Environ Pollut,2016,213:160-172.
[5]林木南,刘献祥.透骨消痛方治疗膝骨性关节炎30例[J].福建中医药,2005,36(4):15-16.
[6]郑春松,朱晓勤,曾建伟,等 HPLC法测定透骨消痛方中芍药苷和迷迭香酸的含量[J].福建中医药大学学报,2011,21(6):37-40.
[7]李翠萍,吴民耀,王宏元.3种半数致死浓度计算方法之比较[J].动物医学进展,2012,33(9):89-92.
[8] 赵劲松,于书霞.利用概率单位法估算EC_(50)的研究[J].生态毒理学报,2010,5(3):420-425.
[9] 王琦,陈瑛,吴忆宁,等.原生动物在环境毒理学研究中的应用[J].中国农学通报,2015,31(5):139-142.
[10]TAMARA V B,MARINA V C,VALERY V K,et al.Applying bioassay methods for ecological assessment of the soils from the brownfield Sites [J/OL].Water,Air,& Soil Pollution,2017,228(9).DOI:10.1007/s11270-017-3521-3.
[11] MARINA K,ANNA D,YURI L,et al.Evaluation of toxicity of nanoclays and graphene oxide in vivo:a Paramecium caudatum study[J].Environmental Science:Nano,2016,3(2):442-452.
[12] HENINE S,ROUABHI R,GASMI S,et al.Oxidative stress status,caspase-3,stromal enzymes and mitochondrial respiration and swelling of Paramecium caudatum in responding to the toxicity of Fe3O4 nanoparticles[J].Toxicol Environ Health Sci,2016,8(2):161-167.
[13] 王正方,齐小花,邹明强,等.三聚氰胺对纤毛类原生动物梨形四膜虫的细胞毒性[J].生态毒理学报,2009,4(1):35-39.
[14]RICHARD M,JAMES W,ANDREW A.Toxicity and applications of internalised magnetite nanoparticles within live paramecium caudatum cells[J].Bio Nano Science,2018,8(1):90-94.
[15]MORGUNOV I G,KARPUKHINA O V,KAMZOLOVA S V,et al.Investigation of the effect of biologically active threo-Ds-isocitric acid on oxidative stress in Paramecium caudatum[J].Prep Biochem Biotechnol,2018,48(1):1-5.
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